【作者】 张健；

【导师】 赵春亭；

【作者基本信息】 青岛大学 ， 内科学， 2015， 硕士

【摘要】 目的探讨血清和骨髓间充质干细胞(mesenchymal stem cell,MSC)表面瘦素及其受体在免疫介导再生障碍性贫血小鼠发病过程中的变化及意义。方法建立免疫介导再生障碍性贫血小鼠模型,分别于建模第0,3,6,9,12,15,18天检测血常规,CD4+T、CD8+T细胞亚群比例,IL-2、IFN-γ、IL-4、IL-5等指标变化,行骨髓活检病理学检查,判断模型成功与否。造模后采用ELISA方法检测血清瘦素及其可溶性瘦素受体、免疫组化法检测骨髓瘦素受体、实时荧光定量PCR方法检测骨髓MSC表面瘦素、瘦素受体和骨髓成脂成骨基因变化。对血清瘦素和瘦素受体与免疫指标、血象指标之间的相关性及骨髓MSC表面瘦素及其受体与成脂成骨基因的相关性进行分析。结果(1)小鼠造模后外周血三系迅速降低;IL-2、IFN-γ水平逐渐升高,IL-4、IL-5、CD4+T/CD8+T比值逐渐下降。骨髓活组织检查示造血组织逐渐被脂肪组织取代。造模成功。(2)血清瘦素逐渐上升、瘦素受体逐渐下降。血清瘦素与Th1相关细胞因子IL-2、IFN-γ呈显著正相关,与Th2相关细胞因子、CD4+T/CD8+T比值及血象呈显著负相关。而瘦素受体与之相反。(3)在微环境中,骨髓MSC表面瘦素逐渐上升、瘦素受体逐渐下降。骨髓脂肪化程度逐渐加重,成脂基因C/EBPα、C/EBPβ、PPARγ表达逐渐升高,成骨基因RUNx2逐渐表达下降。瘦素基因表达与成脂基因表达呈正相关,与成骨基因表达呈负相关。瘦素受体亦与之相反。结论1.再障小鼠模型可模拟人体再障的发病过程;2.瘦素可能加重再障免疫紊乱过程;3.在微环境中存在瘦素受体减少、瘦素失利用现象。更多还原

【Abstract】 Objective To investigate the role of leptin and its receptor in the serum and bone marrow mesenchymal stem cell(MSC) of immune-mediated aplastic anemia mice.Methods Aplastic anemia was induced by immune-mediated method. The blood routine、the proportion of CD4+T、CD8+T cells and serum IL-2、IFN-γ、IL-4 and IL-5 were measured on 0,3,6,9,12,15 and 18 days after operation,bone marrow biopsy was taken to determine whether the model is successful.And then the expression of leptin and its receptor in blood was measured by ELSIA,leptin receptor in bone marrow was detected with immunohistochemistry,and the expression of leptin and its receptor in bone marrow mesenchymal stem cell、lipogenic genes and bone-related genes in bone marrow were measured by RT-PCR.The correlation between leptin、leptin receptor in blood and immunity、blood index and between leptin、leptin receptor in bone marrow MSC and lipogenic、bone-related genes were analysed.Results 1.The blood cells were significantly decreased,serum IL-2 and IFN-γconcentration were increased while IL-4 and IL-5 were decreased,the ratio of CD4+ T /CD8+T was also decreased after infusion,Bone marrow hematopoietic tissue was replaced by adipose tissue.These models were susscessful.2. The level of leptin and leptin receptor showed upward and downward trend separately.Serum leptin had significiant positive correlation with Th1 associated cytokines,negative correlation with Th2 associated cytokines and the ratio of CD4+ T /CD8+T. But serum leptin receptor exactly the opposite.3. In microenvironment, the level of leptin and leptin receptor showed upward and downward trend separately,the expression of C/EBPα、C/EBPβ、PPARγshowed upward while RUNx2 showed downward trend. Leptin had significiant positive correlation with lipogenic genes,negative correlation with bone-related genes.But leptin receptor exactly the opposite.Conclusion 1. Aplastic anemia mouse model can imitate the disease process in human.2.Leptin may aggravate the immune dysfunction. 3. Owing to the damagement of leptin receptor,leptin is achrestic in microenvironment,更多还原